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Molecular marker MboII-40 for indicating and identifying watermelon pulp color and application

A technology of mboii-40 and molecular markers, applied in the field of molecular biology, can solve the problems of small number of molecular markers, low polymorphism, narrow genetic distance of watermelon, etc., and achieve the effect of improving breeding efficiency and shortening the breeding period

Active Publication Date: 2015-06-17
青岛新城胶河源绿色农业发展有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the genetic distance of watermelon is relatively narrow, and the number of available molecular markers published in the literature is small and the polymorphism is low, which seriously restricts the gene mapping of main agronomic traits of watermelon and the development of related molecular markers.

Method used

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  • Molecular marker MboII-40 for indicating and identifying watermelon pulp color and application
  • Molecular marker MboII-40 for indicating and identifying watermelon pulp color and application
  • Molecular marker MboII-40 for indicating and identifying watermelon pulp color and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Example 1 Primer Design and Genomic DNA Extraction

[0037] The F 1 , F 2 The generation group was used as experimental material to verify the presence of lycopene in watermelon pulp by CAPS markers. The result is as figure 1 shown.

[0038] Two watermelon lines, COS and LSW-177, were sequenced using high-throughput sequencing technology, and the cds region sequence of the watermelon lycopene β-cyclase gene was obtained through the NCBI website, and compared with the sequenced material, the obtained The chromosomal interval where the cds region sequence of the watermelon lycopene β-cyclase gene exists, within this interval, the base segment with the SNP mutation site in the sequencing data is mined, and the enzyme digestion information of the SNP site is analyzed to obtain the existence For the sequence of CAPS mutation, CAPS primers were designed on the sequence with CAPS site. The primer length is 18-26bp, the annealing temperature is 56-60°C, and the GC content is...

Embodiment 2

[0039] The acquisition of embodiment 2PCR product

[0040] The obtained CAPS primers and genomic DNA were used for PCR reaction, and the PCR reaction system was shown in Table 1 (20 μl):

[0041] Table 1. PCR reaction system

[0042]

[0043] Touchdown PCR (touchdown PCR, TD-PCR) was used for amplification, and the program was: 94°C pre-denaturation for 7 minutes, 94°C denaturation for 30 s, 60°C annealing for 30 s, 0.5°C lowering per cycle, 72°C extension for 40 s, 30 cycles; 94 Denaturation at ℃ for 30s, annealing at 45℃ for 30s, extension at 72℃ for 40s, 10 cycles; extension at 72℃ for 10min, storage at 4℃.

[0044] Utilize 1% agarose gel electrophoresis to detect the obtained PCR product, and the detection result is as follows figure 1 As shown, LSW-177, COS, F 1 and F 2 A fragment with a size of 618bp was amplified from each plant, which was consistent with the expected fragment size.

Embodiment 3

[0045] Embodiment 3 PCR product is carried out enzyme digestion verification

[0046] According to the restriction endonuclease operation guide of Thermo, use restriction endonuclease to check the enzyme digestion of the obtained PCR product. The enzyme digestion system is shown in Table 2: (15.3 μl)

[0047] Table 2. Enzyme digestion reaction system

[0048]

[0049] Enzyme digested overnight in a water bath at 37°C, and the digested products were detected by 1% agarose electrophoresis, and the detection results were as follows: figure 1 shown. LSW-177 was not cleaved by restriction endonucleases because there was no enzyme cleavage site, and it was still 618bp. Due to the presence of an enzyme cleavage site, COS was cleaved by MboII and fragments of 460 and 158bp were obtained. f 1 Representatives exhibiting co-dominant features include not only a 618bp fragment of LSW-177, but also 460 and 158bp fragments. In the tested 18 strains of F 2 Among the generation single ...

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Abstract

The invention discloses a molecular marker MboII-40 for indicating and identifying watermelon pulp color and application and belongs to the technical field of molecular biology. The fragment size of a PCR product amplified by the molecular marker is 618bp, and the nucleotide sequence is as shown in SEQ ID NO.1. The CAPS molecular marker has universality in different materials and can identify watermelons in the watermelon seedling stage, the operation is simple and convenient, and the result is definite. The molecular marker can be used for molecular marker-assisted selection of high lycopene of watermelon in future, and the breeding work efficiency is improved.

Description

technical field [0001] The invention relates to a molecular marker MboII-40 for predicting and identifying the color of watermelon pulp and its application, belonging to the technical field of molecular biology. Background technique [0002] Molecular marker-assisted selection is a new technology produced with the rapid development of modern molecular biology technology. It can quickly and accurately analyze the genetic composition of individuals at the molecular level, so as to realize direct selection of genotype and carry out molecular breeding. The key to the success of molecular marker-assisted selection lies in whether the obtained molecular markers are linked to the traits to be analyzed and whether they are universal among different materials of the same species. [0003] The color of watermelon pulp is an important indicator of watermelon quality. Watermelon fruit contains lycopene, β-carotene, lutein, zeaxanthin and other pigments. The composition and content of di...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11C12N15/10
CPCC12Q1/6895C12Q2600/13C12Q2600/156
Inventor 刘宏宇朱子成栾非时刘佳俊
Owner 青岛新城胶河源绿色农业发展有限公司
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